Enhanced angiogenic and cardiomyocyte differentiation capacity of epigenetically reprogrammed mouse and human endothelial progenitor cells augments their efficacy for ischemic myocardial repair.
Enhanced angiogenic and cardiomyocyte differentiation capacity of epigenetically reprogrammed mouse and human endothelial progenitor cells augments their efficacy for ischemic myocardial repair.
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DOI：
10.1161/circresaha.112.270462
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发表时间：
2012-07-06
影响因子：
20.1
通讯作者：
Kishore R
中科院分区：
文献类型：
--
作者：
Thal MA;Krishnamurthy P;Mackie AR;Hoxha E;Lambers E;Verma S;Ramirez V;Qin G;Losordo DW;Kishore R
尽管骨髓内皮祖细胞疗法(BM-EPC)改善了缺血性心脏病患者的症状，但其有限的可塑性和现有心脏病患者的功能下降限制了EPC疗法在心脏再生医学中的充分好处。我们假设，使用针对关键表观遗传抑制标记的小分子对小鼠和/或人内皮祖细胞进行重新编程将导致激活基因转录的全球增加，诱导它们的心肌生成潜力，并增强它们固有的血管生成潜力。用DNA甲基转移酶(5-氮杂胞苷)、组蛋白脱乙酰基酶(丙戊酸)和G9a组蛋白二甲基转移酶抑制剂处理小鼠LIN-Sca1+CD31+内皮祖细胞和人CD34+细胞。48h处理后，活性转录组的表达增加，包括多能相关基因的重新激活和CMC特异性基因的表达，而EC特异性基因的表达明显上调。在适当的分化条件下，重编程内皮祖细胞能有效地分化为CMC和血管平滑肌细胞。用表观遗传修饰剂处理后，心肌细胞和多能细胞特异性基因启动子上的组蛋白乙酰化显著增加。在急性心肌梗死小鼠模型中，重新编程的小鼠和人内皮祖细胞心肌内移植显示出显著的心功能改善，这在组织学上得到了其新生CMC分化的支持，并增加了毛细血管密度和减少了纤维化。重要的是，细胞移植是安全的，不会形成畸胎瘤。综上所述，我们的结果表明，表观重编程内皮祖细胞表现出更安全、更可塑性的表型，并通过新的心肌生成和新的血管形成改善梗死后的心脏修复。
While Bone-marrow endothelial progenitor cell based therapies (BM-EPC) improve the symptoms in patients with ischemic heart disease their limited plasticity and decreased function in patients with existing heart disease limits the full benefit of EPC therapy for cardiac regenerative medicine. We hypothesized that reprogramming mouse and/or human EPCs using small molecules targeting key epigenetic repressive marks would lead to a global increase in active gene transcription, induce their cardiomyogenic potential and enhance their inherent angiogenic potential. Mouse Lin-Sca1+CD31+ EPCs and human CD34+ cells were treated with inhibitors of DNA methyltransferases (5-Azacytidine), histone deacetylases (valproic acid) and G9a histone di-methyltransferase. Forty eight hour treatment led to global increase in active transcriptome including the reactivation of pluripotency associated and CMC specific mRNA expression while EC specific genes were significantly up-regulated. When cultured under appropriate differentiation conditions, reprogrammed EPCs showed efficient differentiation into CMC and vascular smooth muscle cells. Treatment with epigenetic modifying agents show marked increase in histone acetylation on cardiomyocyte and pluripotent cell specific gene promoters. Intra-myocardial transplantation of reprogrammed mouse and human EPCs in an acute myocardial infarction mouse model showed significant improvement in ventricular functions, which was histologically supported by their de novo CMC differentiation and increased capillary density and reduced fibrosis. Importantly, cell transplantation was safe and did not form teratomas. Taken together, our results suggest that epigenetically reprogrammed EPCs display a safe, more plastic phenotype and improve post-infarct cardiac repair by both neo-cardiomyogenesis and neovascularization.